Project description:Single cell sequencing in peripheral blood mononuclear cells (PBMCs) revealed a novel human-specific long noncoding RNA called heart-failure associated transcript 4 (HEAT4). HEAT4 expression was assessed in several in vitro and ex vivo models of immune cell activation, as well as in the blood of patients with heart failure (HF), acute myocardial infarction (AMI) and cardiogenic shock (CS). The transcriptional regulation of HEAT4 was verified through cytokine treatment and single cell sequencing. Loss-of-function and gain-of-function studies and multiple RNA–protein interaction assays uncovered a mechanistic role of HEAT4 in the monocyte anti-inflammatory gene program. HEAT4 expression and function was characterized in a vascular injury model in NOD.CB-17-Prkdc scid/Rj mice.
Project description:Rationale: Airspace macrophages are the most abundant cell in airspaces and are viewed as a homogeneous population during health. Single cell RNA sequencing allows for examination of transcriptional heterogeneity between cells and between individuals. Understanding the conserved repertoire of airspace leukocytes during health is essential to understanding cellular programing during disease. Objective: We sought to determine the transcriptional heterogeneity of human bronchoalveolar lavage cells in healthy adults. Methods: Ten healthy subjects underwent bronchoscopy. Cells obtained from lavage fluid were subjected to single cell RNA sequencing. Unique cell populations and putative functions were identified. Transcriptional profiles were compared across individuals. Measurements and Main Results: Based on transcriptional profiling we identify highly conserved macrophage, monocyte-like, lymphocyte, dendritic cell, and cycling cell populations. We define two unique subgroups of resident airspace macrophages - one defined by a pro-inflammatory profile and one by metallothionein gene expression. We identify distinct subsets of monocyte-like cells and directly compared them to peripheral blood mononuclear cells. Finally, we compare global macrophage and monocyte programing between male and female subjects. Conclusions: Healthy human airspaces contain multiple populations of leukocytes that are highly conserved between individuals and between the sexes. Resident macrophages comprise the largest population and include novel subsets defined by inflammatory and metal-binding gene signatures. Monocyte-like cells within the airspaces are transcriptionally distinct from circulating blood cells and include a rare population defined by expression of cell-matrix interaction genes. This study is the first to define airspace immune cell heterogeneity and identifies three previously unrecognized myeloid cell subsets.
Project description:Interventions: Sevoflurane group:Nil;Propofol group:NIl
Primary outcome(s): Heterogeneity of immune cell RNA spectrum in local tissue and peripheral blood of tumor
Study Design: Non randomized control
Project description:Single cell-based studies have revealed tremendous cellular heterogeneity in stem cell and progenitor compartments, suggesting continuous differentiation trajectories with intermixing of cells at various states of lineage commitment and notable degree of plasticity during organogenesis. The hepato-pancreato-biliary organ system relies on a small endoderm progenitor compartment that gives rise to a variety of different adult tissues, including liver, pancreas, gallbladder, and extra-hepatic bile ducts. Experimental manipulation of various developmental signals in the mouse embryo underscored important cellular plasticity in this embryonic territory. This is also reflected in the existence of human genetic syndromes as well as congenital or environmentally-caused human malformations featuring multiorgan phenotypes in liver, pancreas and gallbladder. Nevertheless, the precise lineage hierarchy and succession of events leading to the segregation of an endoderm progenitor compartment into hepatic, biliary, and pancreatic structures are not yet established. Here, we combine computational modelling approaches with genetic lineage tracing to assess the tissue dynamics accompanying the ontogeny of the hepato-pancreato-biliary organ system. We show that a multipotent progenitor domain persists at the border between liver and pancreas, even after pancreatic fate is specified, contributing to the formation of several organ derivatives, including the liver. Moreover, using single-cell RNA sequencing we define a specialized niche that possibly supports such extended cell fate plasticity.
Project description:We used single cell RNA sequencing (scRNA-seq) to characterize the heterogeneity of peripheral blood-derived canine iNKT cells relative to that of human iNKTs.
Project description:The ontogeny of human lung macrophages derived from blood monocytes is poorly understood. In this study, we employed single-cell RNA-sequencing to investigate the heterogeneity of HSPC-derived human lung monocytes and macrophages in the MISTRG humanized mouse model.
Project description:We combined the Single-probe single cell MS(SCMS) experimental technique with a bioinformatics software package, SinCHet-MS (Single Cell Heterogeneity for Mass Spectrometry), to characterize changes of tumor heterogeneity, quantify cell subpopulations, and prioritize the metabolite biomarkers of each subpopulation.
Project description:In atherosclerosis progression and regression, monocytes or monocyte-derived macrophages are the major immune cells in the plaque. It is important to understand the fate and characteristics of monocyte/macrophage during the plaque progression and regression. To characterize the fate of monocytes/macrophages, we performed single cell RNA sequencing of fate-mapped aortic CX3CR1-derived monocytes/macrophages from Cx3cr1CreERT2-IRES-YFP/+Rosa26floxed-tdTomato/+ mice with AAV-PCSK9 injection and fed a Western Diet. The single cell RNA-seq analyses revealed the heterogeneity of aortic macrophages and identified a stem-like cell cluster in atherosclerotic aorta.
Project description:As supplies of monocytes, macrophages and dendritic cells from human sources can be scarce or prone to donor variation we established an efficient method to generate induced pluripotent stem cell derived monocytes that in turn could be differentiated into both macrophages and dendritic cells. We used RNA sequencing to profile these from multiple differentiation runs (n=3) and multiple monocyte harvests (n=3-4) and compared them to their blood derived counterparts, blood derived monocyte, monocyte derived macrophages and moncyte derived dendritic cells (from 3 donors).
Project description:In order to gain deep insights into the molecular characteristics and heterogeneity of MDSCs in human newborns, low-density CD11b+HLA-DR-/low immature myeloid cells from the peripheral blood of term infants, preterm infants, and adults control were subjected to single-cell RNA sequencing (scRNA-seq).